Particle for detecting enzyme activity, method for detecting the enzyme activity and enzyme activity detection device by the use thereof

Abstract

An object is to provide a particle for detecting an enzyme activity wherein by measuring an intensity of a fluorescence wavelength in a sample solution, it is possible to not only detect the presence or absence of an enzyme but also accurately perform a quantitative analysis of the enzyme in the sample solution, handleability is excellent as well as measurement accuracy and quantitative property can be enhanced because the present invention is hardly influenced by absorbed moisture and weighing errors hardly occur, and further productivity is remarkably excellent because complicated steps such as a step of cleaving and purifying a synthesized peptide and a step of lyophilizing are not required upon production thereof. The particle 1 for detecting the enzyme activity of the present invention comprises a particle 2 and a first compound 3 binding the particle 2 to one end, binding a first fluorescent group 4 to the other end and having a cleavage site by an enzyme 5 in a sample solution.

1 . A particle for detecting an enzyme activity comprising a particle and a first compound binding the particle to one end, binding a first fluorescent group to the other end and having a cleavage site by an enzyme. 
   
   
       2 . The particle for detecting the enzyme activity according to  claim 1 , wherein the first fluorescent group is fluorescein or a derivative thereof. 
   
   
       3 . A particle for detecting an enzyme activity comprising a second compound binding a second fluorescent group to one end, binding a quenching agent to an atomic group at the other end and having a cleavage site by an enzyme, and a particle bound to the atomic group or the quenching agent at the other end in the second compound. 
   
   
       4 . The particle for detecting the enzyme activity according to any one of  claims 1  to  3 , wherein the first compound or the second compound has an acetylated amino acid residue. 
   
   
       5 . A method for detecting an enzyme activity comprising (a) a contact reaction step of contacting and reacting a sample solution containing an enzyme with the particle for detecting the enzyme activity according to any one of  claims 1  to  3 , and (b) a fluorescence measurement step of measuring fluorescence in the sample solution reacted with the particle for detecting the enzyme activity. 
   
   
       6 . A method for detecting an enzyme activity comprising (a) a contact reaction step of contacting and reacting the particle for detecting the enzyme activity of  claim 4  with a sample solution containing a deacetylase which liberates an acetyl group from the first compound or the second compound having an acetylated amino acid residue and a particular enzyme which selectively cleaves a peptide bond of the amino acid residue whose acetyl group has been liberated, and (b) a fluorescence measurement step of measuring fluorescence in the sample solution reacted with the particle for detecting the enzyme activity. 
   
   
       7 . An enzyme activity detection device comprising a vessel comprising a liquid inlet formed upstream of a liquid passing path and a liquid outlet formed downstream of the liquid inlet and the particle for detecting the enzyme activity according to any one of  claims 1  to  4  housed in the liquid passing path and retained in the vessel.